This research application focuses on the generation of HIV-1 resistant primary T cells as potential translational immunotherapies for HIV-1. The specific aims include: 1) engineering primary T cells with a membrane bound form of the HIV-1 fusion inhibitor C34, evaluation of antiviral effects and cellular immune function, 2) development of strategies to compare and combine multi-target transgenes in a lentiviral backbone vector and 3) investigation of the immunogenicity of HIV epitopes exposed as a result of binding to the fusion inhibitor expressed on the target cell. A self-inactivating lentiviral vector system will be used to deliver genes coding for membrane bound forms of an HIV-1 entry inhibitor alone, and in combination with small interfering RNAs against HIV-1 co-receptor CCR5, and HIV genes Rev and Tat. Transduced primary T cells will be challenged with a variety of HIV-1 viruses including primary isolates, and cells will be evaluated for immunologic function. Preliminary studies in primary T cells are promising for the membrane bound entry inhibitor, and we hypothesize that combinations with siRNA against viral co-receptor and/or regulatory HIV genes will have an additive anti-viral effect. This proposal describes a 5-year training program for the development of an academic career in Pediatric Allergy and Immunology (A/I). The principal investigator has completed Pediatric residency training at the Children's Hospital of Philadelphia, and is in her third year of A/I Fellowship. This grant will allow the investigator to broaden her scientific skills while benefiting from the diverse resources available at the University of Pennsylvania (UPENN) within the Department of Immunology, the Center for AIDS Research (CFAR), and the Abramson Family Cancer Research Institute (AFCRI). Drs. Carl June and James Hoxie will mentor Dr. Perez's scientific career development. Dr. June is the Director of the Translational Research Program of the AFCRI, and has led pioneering translational research in T cell biology and adoptive immunotherapy of cancer and HIV infection. Dr. Hoxie, Director of the UPENN CFAR, has an extensive background in HIV virology, pathogenesis and viral entry. [unreadable] [unreadable]